Method to specifically stimulate survival and expansion of genetically-modified immune cells

Abstract

The present invention relates to a vector encoding a wildtype or truncated form of erythropoietin receptor (EpoR) to promote T cell survival and proliferation. Specifically, it exemplifies a bicistronic vector that expresses a truncated EpoR with an anti-CD19-41 BB-003ζ chimeric antigen receptor (CAR) that has a greater ex vivo expansion than CAR-T cells and demonstrates a significantly higher anti-leukemic activity in vivo. It also describes the method of producing cells expressing said vector and the use of these cells to kill CD19+ tumour cells.

What is claimed is: 
     
         1 . A mammalian T cell comprising a vector, the vector comprising a nucleic acid encoding:
 a) an erythropoietin (Epo) receptor;   b) a 2A peptide; and   c) a chimeric antigen receptor comprising:
 i) a signal peptide; 
 ii an extracellular receptor domain that binds CD19; 
 iii) a hinge and transmembrane domain that anchors the extracellular receptor domain on the surface of a cell; and 
 iv) an effector domain; 
   wherein the Epo receptor is expressed on the mammalian T cell surface, and   wherein the nucleic acid encodes SEQ ID NO: 12.   
     
     
         2 . A method of making a transgenic mammalian T cell, the method comprising introducing a vector into a mammalian T cell, the vector comprising a nucleic acid encoding:
 a) an erythropoietin (Epo) receptor;   b) a 2A peptide; and   c) a chimeric antigen receptor comprising:
 i) a signal peptide; 
 ii) an extracellular receptor domain that binds CD19; 
 iii) a hinge and transmembrane domain that anchors the extracellular receptor domain on the surface of the mammalian T cell; and 
 iv) an effector domain; 
   
       wherein the Epo receptor is expressed on the mammalian T cell surface, and 
       wherein the nucleic acid encodes SEQ ID NO: 12. 
     
     
         3 . The method of  claim 2 , wherein the mammalian T cell further expresses a T-cell receptor (TCR) that binds a tumor antigen or a viral antigen. 
     
     
         4 . The method of  claim 3 , wherein the TCR is endogenous. 
     
     
         5 . The method of  claim 4 , wherein the mammalian T cell is a tumor-infiltrating lymphocyte (TIL), and wherein the method further comprises extracting the tumor-infiltrating lymphocyte from a tumor and expanding the TIL ex vivo. 
     
     
         6 . The mammalian T cell of  claim 1 , wherein the mammalian T cell is a human T cell. 
     
     
         7 . The mammalian T cell of  claim 1 , wherein the mammalian T cell is a human peripheral blood T lymphocyte. 
     
     
         8 . The mammalian T cell of  claim 1 , wherein the mammalian T cell expresses a T-cell receptor (TCR) that binds a tumor antigen or a viral antigen. 
     
     
         9 . The mammalian T cell of  claim 8 , wherein the TCR is endogenous. 
     
     
         10 . The mammalian T cell of  claim 9 , wherein the mammalian T cell is a tumor-infiltrating lymphocyte (TIL). 
     
     
         11 . The mammalian T cell of  claim 1 , wherein the mammalian T cell is a CD8+ T cell. 
     
     
         12 . The method of  claim 2 , wherein the mammalian T cell is a CD8+ T cell.